Sperm morphology analysis using strict criteria as a prognostic factor in intrauterine insemination

The objective of this study was to investigate the predictive value of Kruger's criteria for sperm morphology on intrauterine insemination (IUI) outcome. A total of 209 infertile patients underwent 244 IUI treatment cycles. These include 75 couples (80 cycles) with teratozoospermia and 134 couples (164 cycles) with unexplained infertility. The pregnancy rates per IUI cycle were 3.8 (1/26), 18.5 (10/54) and 29.9% (49/164) in patients with sperm morphology with <4, 4-9 and >9% normal forms, respectively, according to Kruger's criteria. A statistical difference in outcome was seen between couples with <4 and >9% normal forms (p = 0.005). Although the difference in pregnancy rates between those with 4-9 and <4% normal forms was not statistically significant, the pregnancy rate for those with 4-9% normal forms was acceptable and still higher than in those with <4% normal forms. Therefore, we suggest that IUI is a reasonable first-line therapy for patients with sperm morphology >4% normal forms, while couples with <4% normal forms should be advised to use in vitro fertilization with intracytoplasmic sperm injection instead of IUI.     

Cationic liposome-mediated GDNF gene transfer after spinal cord injury

Glial cell line-derived neurotrophic factor (GDNF) has been shown to protect cranial and spinal motoneurons, which suggests potential uses of GDNF in the treatment of spinal cord injury (SCI) and motor neuron disease. We examined neuroprotective effect of cationic liposome-mediated GDNF gene transfer in vivo on axonal regeneration and locomotor function recovery after SCI in adult rats. The mixture of DC-Chol liposomes and recombinant plasmid pEGFP-GDNF cDNA was injected after SCI. RT-PCR confirmed the increased expression of GDNF mRNA in the injected areas at 7 days after injection. The expression of EGFP-GDNF was observed in the cells around the injection locus by fluorescence microscope at least 4 weeks after injection. Four weeks after GDNF gene transfer, regeneration of the corticospinal tracts was assessed using anterograde tract tracing. There are more HRP labeling of corticospinal tract axons across the lesion in GDNF group compared with control group. In GDNF group, the maximum distance these labeled axons extended varied in different animals and ranged from 5 mm to approximately 9 mm from the lesion. In control group, no HRP labeled axons extended caudal to the lesion. The locomotion function of hindlimbs of rats was evaluated using inclined plane test and BBB locomotor scores. The locomotion functional scores in GDNF group were higher than that in control group within 1-4 weeks after SCI (p < 0.05). These data demonstrate that in vivo transfer of GDNF cDNA can promote axonal regeneration and enhance locomotion functional recovery, suggesting that cationic liposome-mediated delivery of GDNF cDNA may be a practical gene transfer method for traumatic SCI treatment.     

Platelet-activating factor in vasoobliteration of oxygen-induced retinopathy

PURPOSE: To test whether platelet-activating factor (PAF) directly causes retinovascular endothelial cell (EC) death. METHODS: Retinovascular density was calculated in rat pups exposed to 80% O(2) from postnatal days (P)6 to P14 (to produce oxygen-induced retinopathy [OIR]), using the adenosine diphosphatase (ADPase) technique, in animals treated with distinct PAF receptor blockers (PCA-4248, BN52021, or THG315). PAF levels were then measured in the retinas. Viability of ECs from piglets and humans in response to C-PAF (a stable PAF analogue) was determined by the reduction of the tetrazolium salt 3-(4,5-dimethyl thiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) by viable cells, incorporation of propidium iodide (PI), TUNEL assay, and release of lactate dehydrogenase. Release of thromboxane (TX) was measured in the cell media. RESULTS: PAF levels in retina were markedly increased by exposure of isolated rat retinas to H(2)O(2) (1 micro M) and of rat pups placed in 80% O(2). Exposure to 80% O(2) induced retinal vasoobliteration, which was equally significantly inhibited ( approximately 60%) by all PAF receptor blockers tested. C-PAF increased incorporation of PI by isolated rat retinal microvasculature. Also, C-PAF caused time- and concentration-dependent death of cultured retinal ECs, which was prevented by the PAF receptor antagonist CV-3988. This effect of C-PAF was selective on retinal and neurovascular ECs, but not on other ECs. DNA fragmentation (TUNEL) was hardly detected, and inhibition of apoptosis-related processes by nicotinamide, cyclosporin A, and Z-DEVD-FMK and Z-VAD-FMK (caspase inhibitors) barely protected against death in EC, whereas C-PAF increased release of lactate dehydrogenase, implying that necrosis is the nature of EC death. Finally, C-PAF-induced cell death was preceded by an increase in TXB(2) levels and was prevented by TXA(2) synthase inhibition (with CGS12970). CONCLUSIONS: The data suggest PAF plays a major role in vasoobliteration in OIR by triggering death of neuroretinal microvascular ECs. The cell death seems to be mediated at least in part by TXA(2). These effects of PAF may participate in ischemic retinopathies such as diabetes and retinopathy of prematurity.     

New isopimarane diterpene and new cineole type glucoside from Nepeta prattii

Together with sixteen known compounds, a new isopimarane diterpene (prattol) and a new cineole type glucoside were isolated from Nepeta prattii. Their structures were elucidated on the basis of spectral methods as isopimar-15-en-3 beta,8 beta,20-triol, and (1R, 2R, 4S)-1,8-epoxy-p-methan-2-O-beta-D-glucopyranosyl(1-->6)-beta- D-glucopyranoside.     

反式曲马朵及其活性代谢产物反式氧去甲基曲马朵肾脏清除的立体选择性

目的:研究反式曲马朵及其活性代谢物反式氧去甲基曲马朵肾脏清除的立体选择性.方法:取雄性SD大鼠的右肾,分别以含反式曲马朵(300μg/L)或反式氧去甲基曲马朵(50μg/L)的灌流液(10mL)进行灌流;利用高效毛细管电泳法测定灌流后灌流液、尿液中反式曲马朵及反式氧去甲基曲马朵对映体的浓度,并计算对映体浓度比值.结果:以反式曲马朵进行离体肾灌流后,在灌流液中,( )-反式曲马朵的浓度高于(-)-反式曲马朵的浓度,( )-反式氧去甲基曲马朵的浓度低于(-)-反式氧去甲基曲马朵的浓度;在尿液中,( )-反式曲马朵较(-)-反式曲马朵多,( )-反式氧去甲基曲马朵较(-)-反式氧去甲基曲马朵少.以反式氧去甲基曲马朵灌流后,在灌流液中( )-反式氧去甲基曲马朵的浓度低于(-)-反式氧去甲基曲马朵的浓度;在尿液中( )-反式氧去甲基曲马朵较(-)-反式氧去甲基曲马朵高.结论:反式曲马朵及反式氧去甲基曲马朵的肾脏清除具有立体选择性.在肾脏中反式曲马朵的氧去甲基代谢具有立体选择性,以(-)-反式曲马朵优先代谢成(-)-反式氧去甲基曲马朵.反式氧去甲基曲马朵的尿排泄具有立体选择性,以( )-反式氧去甲基曲马朵占优.     

Effects of B7-blocking agent and/or CsA on induction of platelet-specific T-cell anergy in chronic autoimmune thrombocytopenic purpura

Chronic autoimmune thrombocytopenic purpura (AITP) is characterized by platelet-specific autoantibody production that is influenced by enhanced antiplatelet T-helper cell reactivity. Costimulatory signals are absolutely required for T-cell activation and play key roles in the decision between tolerance and immunity. In this study we cultured T cells isolated from patients with chronic AITP to investigate the effects of the B7-blocking agent cytologic T-lymphocyte-associated antigen 4-immunoglobulin (CTLA4-Ig), and cyclosporin A (CsA), alone or in combination, on induction of platelet-specific T-cell anergy. The data showed that in most cases CTLA4-Ig and/or CsA could induce tolerance toward platelet antigens based on anergy. It could be overcome by stimulation with unrelated antigens, demonstrating its platelet specificity. The anergy is associated with lack of interleukin 2 (IL-2) and withheld by exogenous IL-2, emphasizing the pivotal role of IL-2 suppression in the induction of platelet-specific anergy. We also prospectively evaluated the efficacy of CsA therapy in patients with refractory AITP and observed that the response to CsA treatment in vivo was associated with the inhibiting sensitivity of platelet-reactive T cells to CsA in vitro. This suggests that the sensitivity of T cells to CsA in vitro could serve as a reliable parameter in predicting the efficacy of CsA for patients with refractory AITP. CTLA4-Ig may become a promising new therapeutic agent for the treatment of chronic AITP, and the combination of CTLA4-Ig and CsA would be a more powerful strategy for the management of refractory AITP.     

Alteration of gap junctions and connexins in the right atrial appendage during cardiopulmonary bypass

OBJECTIVES: We investigated the influence of cardiopulmonary bypass on cardiomyocyte gap junctions and connexins. METHODS: Samples were collected at intervals during operation from the right atrial appendage in 21 patients (mean [+/- SD] age 55 +/- 21 years). Immunodetection of connexins was conducted by Western blotting and confocal microscopy with parallel electron microscopic examination of gap junctions. RESULTS: Downregulation of connexin 43 during the course of operation occurred in more than half of the patients. The mean densitometric value of connexin 43 decreased by 23%, with samples from patients with coronary artery disease showing a greater reduction than seen in those from patients with other diseases (31% +/- 22% vs 10% +/- 24%, P =.04). Such alterations were confirmed by confocal microscopy, which also demonstrated reduced connexin 45 immunolabeling in most patients. Electron microscopy revealed a reduction in the dimensions of cell membrane-located gap junctions and more frequent intracytoplasmic gap junctional membrane in samples from later time points (P =.04). CONCLUSIONS: Downregulation of connexins accompanied by a reduction in gap junctions is common in the cardiomyocytes of the right atrial appendage during cardiopulmonary bypass. The association of a marked reduction in connexin 43 with coronary artery disease may imply inadequate intraoperative cardiac protection in patients with this disease.